The Bric-a-brac, Tramtrack, Broad-complex (BTB) domain is a protein-protein interaction domain that is found in many zinc finger transcription factors. BTB containing proteins play important roles in a variety of cellular functions including regulation of transcription, regulation of the cytoskeleton, protein ubiquitination, angiogenesis, and apoptosis. Here, we report the cloning and characterization of a novel human gene, KLHL31, from a human embryonic heart cDNA library. The cDNA of KLHL31 is 5743 bp long, encoding a protein product of 634 amino acids containing a BTB domain. The protein is highly conserved across different species. Western blot analysis indicates that the KLHL31 protein is abundantly expressed in both embryonic skeletal and heart tissue. In COS-7 cells, KLHL31 proteins are localized to both the nucleus and the cytoplasm. In primary cultures of nascent mouse cardiomyocytes, the majority of endogenous KLHL31 proteins are localized to the cytoplasm. KLHL31 acts as a transcription repressor when fused to GAL4 DNA-binding domain and deletion analysis indicates that the BTB domain is the main region responsible for this repression. Overexpression of KLHL31 in COS-7 cells inhibits the transcriptional activities of both the TPA-response element (TRE) and serum response element (SRE). KLHL31 also significantly reduces JNK activation leading to decreased phosphorylation and protein levels of the JNK target c-Jun in both COS-7 and Hela cells. These results suggest that KLHL31 protein may act as a new transcriptional repressor in MAPK/JNK signaling pathway to regulate cellular functions.
Fish sauces prepared from sardine, Sardinops melanosticta were tested for inhibitory activity against angiotensin-I converting enzyme(ACE). Three kinds of fish sauces were prepared from scrap(S), meat(M) and round(R) of sardine, respectively. ACE inhibitory activity of sardine sauce S and R decreased with the elapse of fermentation period, whereas that of sardine sauce M increased to 30 days and thereafter decreased. ACE inhibitory activity of sardine sauce M fermented with koji was higher than that without koji. And occurrence of $5\%$ TCA soluble peptide-nitrogen was similar to tendancy of the ACE inhibitory activity. The ACE inhibitory activity increased with an increment of amounts added and was stable at heat treatment in boiling water bath for 5hrs. $IC_{50}\%$ (Amounts of inhibitors need for $50\%$ inhibition) of the sardine sauce S, M and R fermented with(without) koji during 90 days was $125{\mu}g(140{\mu}g),\;200{\mu}g(100{\mu}g)$ and $125{\mu}g(135{\mu}g)$, respectively. From the profiles of fractionation of the sardine sauce R fermented without koji for 90 days, the molecular weight of most active fraction was about 1,400 and the amino acids of Glu, Ala, Leu and Lys were found in abundance.
For the Exp. 1, a total of sixty pigs(15.95${\pm}$0.09kg average initial body weight) were used in a 28-d growth assay to determine the effects of extruded chinese corn on growth performance and nutrient digestibility in nursery pigs. Dietary treatments included 1) UCORN(U.S. corn-SBM based diet), 2) CCORN(Chinese corn-SBM based diet) and 3) ECCORN(Extruded Chinese corn-SBM based diet). Overall period, average daily gain of pigs fed ECCORN diet was higher than that of pigs fed CCORN diet(547 vs 522 g/d), however, there was not significant difference. On day 10 of the experiment, pigs fed UCORN and ECCORN diet had significantly increased in DM and DE digestibilities compared to pigs fed CCORN diet(P〈0.05). Also, on day 24 of the experiment, pigs fed UCORN and ECCORN diet had a significant increase in DM digestibility compared to pigs fed CCORN diet(P〈0.05). Pigs fed ECCORN diet had significantly increased DE digestibility compared to pigs fed CCORN diet(P〈0.05). For the Exp. 2, three cannulated barrows(54.09kg average initial body weight) were used to determine the apparent ileal digestibilities of amino acids and nutrient digestibility of extruded corn in finishing pigs. Dietary treatments were the same as in Exp. 1. Apparent ileal digestibility of cystine was greater for UCORN and ECCORN than for CCORN(P〈0.05). Apparent digestibility of DM at the total tract was greater for UCORN and ECCORN than for CCORN(P〈0.05). Pigs fed UCORN and ECCORN diet had a significant increase in apparent total tract digestibility of N compared to pigs fed CCORN diet(P〈0.05). In conclusion, the results obtained from these feeding trials suggest that the extruded corn for nursery pigs had affected growth performance and DM and DE digestibilities. In finishing pigs, extruded corn was an effective means to improve apparent total tract digestibilities of DM and N.
The nutritional properties of the Chol-Pyon were investigated with changing the materials (mugwort and pine leaves). In proximate composition, rice powder added mugwort and pine leaves showed the lligher con-tents of crude protein, crude lipid and crude ash than in rice powder. Ihe pH of rice powder, mugwort and pine leaves was 6.4, 6.8 and 3.5, respectively. The rice powder added pine leaves showed the lowest pH value. The content of the free sugar in raw materials for ChOl-PyOn preparation was 0.9% in rice powder, 0.3% in mugwort and 2.7% in pine leaves. Eighteen kinds of amino acids were determined in raw materials for ChOl-fyOn preparation and their contents were 4.8% in mugwort, 4.2% in rice powder and 2.8% in pine leaves. The major minerals of raw materials for ChOl-PyOn preparation was 0.9% increased in the order of K> Na > Mg > Ca in rice powder, Mg > K > Ca > Na in mugwort, and K > Ca > Mg > Na in pine leaves. Both of mugwort and pine leaves additives showed the higher contents of 8 kinds of minerals (Ca, Mg, K, Na, Mn, Fe, Cu, Zn) than in rice powder. In relation to changes in the texture of ChOl-PyOn, hardness, fracturability and adhesiveness at 25${\pm}$1$^{\circ}C$ were measured to be highest in white ChOl-PyOn. Cohesiveness was shown to be highest at 15% in case of mugwort and 2.5% in case of pine leaves. Elasticity was measured to be highest at 0.99 in case that 7.5% mugwort was added to raw materials for ChOl-PyOn. As a result of estimating the sensory qualities of the ChOl-PyOn prepared to which the additives were added in differing amounts, immediately after its preparation the mugwort additive of 7.5% showed the superior sensory qualities Chol-PyOn (p < 0,01).
We established a high throughput screening system of African yam tuber lines which contain high contents of total carotenoids, flavonoids, and phenolic compounds using ultraviolet-visible (UV-VIS) spectroscopy and Fourier transform infrared (FT-IR) spectroscopy in combination with multivariate analysis. The total carotenoids contents from 62 African yam tubers varied from 0.01 to $0.91{\mu}g{\cdot}g^{-1}$ dry weight (wt). The total flavonoids and phenolic compounds also varied from 12.9 to $229{\mu}g{\cdot}g^{-1}$ and from 0.29 to $5.2mg{\cdot}g^{-1}$dry wt. FT-IR spectra confirmed typical spectral differences between the frequency regions of 1,700-1,500, 1,500-1,300 and $1,100-950cm^{-1}$, respectively. These spectral regions were reflecting the quantitative and qualitative variations of amide I, II from amino acids and proteins ($1,700-1,500cm^{-1}$), phosphodiester groups from nucleic acid and phospholipid ($1,500-1,300cm^{-1}$) and carbohydrate compounds ($1,100-950cm^{-1}$). Principal component analysis (PCA) and subsequent partial least square-discriminant analysis (PLS-DA) were able to discriminate the 62 African yam tuber lines into three separate clusters corresponding to their taxonomic relationship. The quantitative prediction modeling of total carotenoids, flavonoids, and phenolic compounds from African yam tuber lines were established using partial least square regression algorithm from FT-IR spectra. The regression coefficients ($R^2$) between predicted values and estimated values of total carotenoids, flavonoids and phenolic compounds were 0.83, 0.86, and 0.72, respectively. These results showed that quantitative predictions of total carotenoids, flavonoids, and phenolic compounds were possible from FT-IR spectra of African yam tuber lines with higher accuracy. Therefore we suggested that quantitative prediction system established in this study could be applied as a rapid selection tool for high yielding African yam lines.
Epidermal growth factor(EGF), a single chain polypeptide of 53 amino acids with a molecular weight of 6,045 Da, was first isolated from the male mouse submandibular glands. EGF stimulates cellular proliferation and differentiation in several tissues and accelerates the rate of wound healing. EGF is bound to the specific receptor(EGFR) on the cell membrane of its target cell. EGFR is a transmembrane glycoprotein with a molecular weight of 170,000 Da and is detectable on a large variety of cell types and tissues. The authors investigated the expression of EGFR in the normal and inflamed human gingival epithelium to study the role of EGFR in the inflammation of the gingival epithelium, and the expression of EGFR in the dental follicle by using in situ mRNA hybridization and immunohistochenistry. The results weree as follows : 1. The expression of EGFR mRNA in the normal gingival epithelium on in situ mRNA hybridization was mainly localized on the basal cell layer, and the spinous layer was weakly positive The granular and cornified layers were negative 2. The expression of EGFR protein in the normal gingival epithelium on inmunohistochemistry was localized on the cornified and granular layers, and the spinous layer was weakly positive. The basal cell layer was completely negative 3. The expression of EGFR mRNA in the inflamed gingival epithelium on in situ mRNA hybridization was evenly and homogeneously distributed in the whole layers of the gingival epithelium except the cornified layer. The staining intensity appeared to increase progressively from the basal cell layer to the cornified layer. 4. The expression of EGFR protein in the inflamed gingival epithelium on immunohistochemistry was evenly and homogeneously distributed in the whole layers of the gingival epithelium. The staining intensity appeared to increase progressively from the cornified layer to the basal cell layer. 5. Strong positive reaction was seen in the epithelial cell rests of Malassez, whereas only background staining was seen in other cells of the dental follicle. In conclusion, the up-regulation of EGFR in the inflamed gingival epithelium and the high amounts of EGFR in the epthelial cell rests of Malassez in the dental follicle can be regarded as responses to the possible damages to the oral environment to maintain the homeostatic conditions.
Jeon, Myong Je;Jang, Min Kyung;Lee, Sol Jee;Park, Sung Hwan;Kim, Mihyang;Sohn, Jae Hak;Lee, Han-Seung;Lee, Dong-Geun;Lee, Sang-Hyeon
Journal of Life Science
/
v.23
no.6
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pp.796-803
/
2013
Property changes and bacterial characterizations by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) were investigated during the fermentation of Makgeollies by 5 isolated yeast strains. Changes of pH were large between day 0 (pH 6) and day 2 (pH 3) and showed less variation after then. ANOVA analyses revealed that pHs were statistically different with fermentation times (p<0.001), while strains (p=0.60) did not. Acidities were changed from 0.19 to 1.04% and showed rather high increase from day 2, and fermentation times (p<0.001) and strains (p=0.006) represented statistical differences. All strains showed less than 0.150% at amino-type nitrogen contents except S strain showed 0.442% at day 8, and there were no statistical differences with fermentation times (p=0.4558) and strains (p=0.3513). Saccharinities of C strain were higher from day 4, and fermentation times (p<0.0001) and strains (p=0.007) showed statistical differences. Large variation of alcohol concentrations (%) were observed between day 0 (0%) and day 2 (10%) and showed less variation after day 2, and there was no statistical difference with strains. Dominant prokaryotes were Lactobacillus fermentum and Pediococcus pentosaceus, which producing acids and functional materials. Dominant eukaryote was Saccharomyces cerevisiae, which might be resulted from addition of yeasts.
Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.
Je, Hae-Soo;Kang, Kyung-Hun;Park, Si-Young;Choi, Byeong-Dae;Kang, Young-Mi;Kim, Jeong-Gyun
Korean Journal of Food Science and Technology
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v.48
no.2
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pp.147-152
/
2016
This study was conducted to investigate the physicochemical quality properties and provide basic data for the activation of traditional Kamju of juice type product prepared by mixing malt and extruded rice collet powder. Malt extracts were prepared by extracting the mixture of malt and water at a weight ratio of 25:75 after soaking for 2 h at $45^{\circ}C$. Rice collet powder was prepared by adjusting the barrel temperature to $95^{\circ}C$, screw speed to $3.07{\times}g$, discharge port diameter to 7 mm and a raw material input to 50 kg/h, the powder was then ground to a particle size of 80 mesh. The physicochemical characteristics (pH, color, viscosity, reducing sugars, number of viable cells, free amino acids) and sensory evaluations were conducted at various time points during the saccharification and at different mixing ratios of the extruded rice collet powder to malt extract (5:95, 15:85, 25:75, 35:65, each at $55^{\circ}C$ for 9 h). As a result, with an increase in the proportion of the extruded rice collet powder and saccharification time, the physicochemical properties of traditional Kamju significantly improved (p<0.05). A mixing ratio of 35:65 rice collet powder to malt extract and a saccharification time of 9 h were found to be the most desirable conditions. However, based on the sensory evaluation, a mixing ratio of rice collet powder and malt extract of 25:75 and a saccharification time of 5 h resulted in the most preferable palatability of traditional Kamju (p<0.05). Therefore, the mixing ratio and saccharification time should be determined to provide a better choice with respect to the taste and economic aspects of traditional Kamju.
Journal of the Korean Society of Food Science and Nutrition
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v.46
no.8
/
pp.1012-1017
/
2017
Mesembryanthemum crystallinum (Family: Aizoaceae) is an annual plant consisting of ice crystal-shaped bladder cells, which is responsible for its common name ice plant. This study investigated biological activities according to general components and extraction solvent in order to examine the functionality of ice plant. The total content of free amino acids was 32.57 mg/g, including 4.64 mg/g of L-alanine as the most abundant and 2.60 mg/g of ${\gamma}$-aminobutyric acid. Regarding angiotensin converting enzyme inhibitory activities of solvent fractions of ice plant, ethyl acetate fraction and chloroform fraction showed activities of $33.17{\pm}3.20{\sim}88.19{\pm}3.20%$ and $23.72{\pm}2.89{\sim}86.78{\pm}2.24%$, respectively, similar to $Captopril^{(R)}$ ($19.51{\pm}3.44{\sim}84.72{\pm}1.06%$) and $Enalapril^{(R)}$ ($24.93{\pm}1.12{\sim}91.32{\pm}3.62%$) as positive control groups. Regarding inhibition of lipid droplet production in 3T3-L1 preadipocytes by ice plant, anti-adipogenic activities were $53.00{\pm}0.45{\sim}65.75{\pm}0.31%$ and $44.16{\pm}0.29{\sim}63.32{\pm}0.36%$ in the ethyl acetate fraction and butanol fraction, respectively, showing the lowest lipid droplet production. The chloroform fraction and hexane fraction showed activities of $38.33{\pm}0.09{\sim}56.55{\pm}0.50%$ and $31.17{\pm}0.50{\sim}55.10{\pm}1.93%$, respectively, whereas the water fraction showed activity of $26.32{\pm}2.27{\sim}49.48{\pm}0.05%$. Therefore, all solvent fractions inhibited fat accumulation of 3T3-L1 preadipocytes according to treatment concentration. According to the results above, it would be possible to utilize ice plant as a new health functional material.
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